Several new areas of development and application of the dynamic nmr (nuclear magnetic resonance) probe of the active site of enzymes will be explored. We hope to provide a novel experimental methods for probing the struture of the transition state enzyme complex by studying the nmr spectra of transition state analogs as they reversibly bind to the enzyme. By also applying these techniques to substrates and intermediates stabilized at low temperatures, it should be possible to achieve a detailed understanding of the enzymatic mechanism of action. In addition an nmr study of covalent substrate intermediates will be presented. This should provide answers as to whether a covalent intermediate does, in fact, exist and, most importantly, define the amino acid residue which covalently links to the substrate. These nmr techniques will be applied to the study of ribonuclease A, alpha-chymotrypsin, 3',5'-cyclic-AMP dependent protein kinase, phosphoglycerate mutase and UDP, Galactose epimerase. However, broad application of these ideas to other enzymes are anticipated as well. In addition, our recently proposed 31P chemical shift-torsional angle hypothesis will provide an important new probe of nucleic acids structure. We hope to study the conformational flexibility about the phosphate ester backbone of oligonucleotides and polynucleotides (t-RNA, 5S RNa) by 31P nmr spectroscopy. We plan to probe with this technique subtle polynucleic acid, drug, protein, and carcinogen interactions.